GSM8047982: E155_KO; Mus musculus; RNA-Seq
The lower incisor germs were carefully separated under a stereomicroscope using micro forceps and transferred into the pre-cooled PBS solution. Lower incisor germ cells from Six1 knockout mice at E15.5 and wild-type incisor germ cells at E14.5, E15.5, and E16.5 were obtained for scRNA-seq. Tissues were dissociated using 0.25% trypsin-EDTA at 37°C for 10 minutes, and the digested cells were passed through a 40 μm sterile filter to obtain the filtrate. The dissociated cells were resuspended in 0.2 ml of 1×DPBS and stained with trypan blue for cell viability counting. The scRNA-seq library was constructed using the micro-well-based GEXSCOPE® Single Cell RNA Library Kit. The cell suspension was first diluted to 1.5×105-3.5×105 cells/ml with pre-chilled PBS for microwell chip experiments. Barcoding beads are subsequently collected from the microwell chip, followed by reverse transcription of the mRNA captured by the barcoding beads to obtain cDNA, and PCR amplification. The amplified cDNA is then fragmented and ligated with sequencing adapters. Sequencing was performed on the Illumina NovaSeq 6000 sequencing platform, generating paired-end reads with a length of 150bp.